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Among various available 3D bioprinting techniques, extrusion-based three-dimensional (3D) bioprinting allows the deposition of cell-laden bioink, ensuring predefined scaffold architecture that may offer living tissue regeneration. With a combination of unique characteristics such as biocompatibility, less cell toxicity, and high water content, natural hydrogels are a great candidate for bioink formulation for the extrusion-based 3D bioprinting process. However, due to its low mechanical integrity, hydrogel faces a common challenge in maintaining structural integrity. To tackle this challenge, the rheological properties, specifically the shear thinning behavior (reduction of viscosity with increasing the applied load/shear rate on hydrogels) of a set of hybrid hydrogels composed of cellulose-derived nanofiber (TEMPO-mediated nano-fibrillated cellulose, TO-NFC), carboxymethyl cellulose (CMC), and commonly used alginate, were explored. A total of 46 compositions were prepared using higher (0.5% and 1.0%) and lower percentages (0.005% and 0.01%) of TO-NFC, 1–4% of CMC, and 1–4% of alginate to analyze the shear thinning factors such as the values of n and K, which were determined for each composition from the flow diagram and co-related with the 3D printability. The ability to tune shear thinning factors with various ratios of a nanofiber can help achieve a 3D bio-printed scaffold with defined scaffold architecture. 

Abstract Three-dimensional bioprinting is a promising field in regenerating patient-specific tissues and organs due to its inherent capability of releasing biocompatible materials encapsulating living cells in a predefined location. Due to the diverse characteristics of tissues and organs in terms of microstructures and cell types, a multinozzle extrusion-based 3D bioprinting system has gained popularity. The investigations on interactions between various biomaterials and cell-to-material can provide relevant information about the scaffold geometry, cell viability, and proliferation. Natural hydrogels are frequently used in bioprinting materials because of their high-water content and biocompatibility. However, the dominancy of liquid characteristics of only-hydrogel materials makes the printing process challenging. Polycaprolactone (PCL) is the most frequently used synthetic biopolymer. It can provide mechanical integrity to achieve dimensionally accurate fabricated scaffolds, especially for hard tissues such as bone and cartilage scaffolds. In this paper, we explored various multimaterial bioprinting strategies with our recently proposed bio-inks and PCL intending to achieve dimensional accuracy and mechanical aspects. Various strategies were followed to coprint natural and synthetic biopolymers and interactions were analyzed between them. Printability of pure PCL with various molecular weights was optimized with respect to different process parameters such as nozzle temperature, printing pressure, printing speed, porosity, and bed temperature to coprint with natural hydrogels. The relationship between the rheological properties and shape fidelity of natural polymers was investigated with a set of printing strategies during coprinting with PCL. The successful application of this research can help achieve dimensionally accurate scaffolds. 

Despite being a very popular topic and researched by several scientists, the entire 3D bioprinting process is still subjected to several challenges like geometric fidelity, mechanical complexities, cell viability, and proliferation. Rheological investigations along with the proper design of experiments help to explore the physical and mechanical properties of biomaterials and 3D printed scaffolds that are directly associated with their geometric fidelity. To ensure post-printed structural integrity, viscosity thickeners and crosslinkers were used in this research. Mixtures of Carboxymethyl Cellulose (CMC, viscosity enhancer), Alginate, and CaCl2 and CaSO4 (crosslinkers) were prepared at various concentrations maintaining minimum solid content. For each composition, a set of rheological tests was performed in form of flow, thixotropic, amplitude, and frequency tests. This research presents an overview of controlling the rheological properties of various bio-inks that are viscosity enhancer and pre-crosslinkers dependent, which opens doors to looking at 3D bioprinting in a very different way. 

Three-dimensional bio-printing is a rapidly growing field attempting to recreate functional tissues for medical and pharmaceutical purposes. Development of functional tissues and organs requires the ability to achieve large full-scale scaffolds that mimic human organs. It is difficult to achieve large scaffolds that can support themselves without damaging printed cells in the process. The high viscosity needed to support large prints requires high amounts of pressure that diminishes cell viability and proliferation. By working with the rheological, mechanical, and microstructural properties of different compositions, a set of biomaterial compositions was identified to have high structural integrity and shape fidelity without needing a harmful amount of pressure to extrude. Various large scale-scaffolds were fabricated (up to 3.0 cm, 74 layers) using those hybrid hydrogels ensuring geometric fidelity. This effort can ensure to fabricate large scaffolds using 3D bio-printing processes ensuring proper internal and external geometries. 

Abstract Bioprinting for regenerative medicine has been gaining a lot of popularity in today's world. Despite being one of the rigorously studied fields, there are still several challenges yet to be solved. Geometric fidelity and mechanical complexities stand as roadblocks when it comes to the printability of the customized constructs. Exploring the rheological properties of the compositions helps us understand the physical and mechanical properties of the biomaterials which are closely tied to the printability of the filament and eventually, geometric fidelity of the constructs. To ensure the structural integrity of the constructs, viscosity enhancers such as carboxymethyl cellulose (CMC) and crosslinkers like CaCl2 and CaSO4 were used. These crosslinkers can be used before (precrosslinking) and after (postcrosslinking) the extrusion of considered compositions to investigate and compare the outcome. To do this, mixtures of CMC (viscosity enhancer), Alginate, and CaCl2 and CaSO4 (crosslinkers) were prepared at various concentrations maintaining minimum solid content (≤8%). Each composition was subjected to a set of rheological tests like flow curve for shear thinning behavior, three points thixotropic for recovery rate, and amplitude test for gelation point. Various geometric fidelity identification tests were conducted and correlated with their physical properties. Some compositions were used to fabricate large-scale constructs (in cm-scale) to demonstrate their capability. This research is a thorough investigation of compositions when they are introduced to crosslinkers and viscosity enhancers which can be crucial for the 3D printing world. 

Maintaining shape fidelity of 3D bio-printed scaffolds with soft biomaterials is an ongoing challenge. Here, a rheological investigation focusing on identifying useful physical and mechanical properties directly related to the geometric fidelity of 3D bio-printed scaffolds is presented. To ensure during- and post-printing shape fidelity of the scaffolds, various percentages of Carboxymethyl Cellulose (CMC) (viscosity enhancer) and different calcium salts (CaCl2 and CaSO4, physical cross-linkers) were mixed into alginate before extrusion to realize shape fidelity. The overall solid content of Alginate-Carboxymethyl Cellulose (CMC) was limited to 6%. A set of rheological tests, e.g., flow curves, amplitude tests, and three interval thixotropic tests, were performed to identify and compare the shear-thinning capacity, gelation points, and recovery rate of various compositions. The geometrical fidelity of the fabricated scaffolds was defined by printability and collapse tests. The effect of using multiple cross-linkers simultaneously was assessed. Various large-scale scaffolds were fabricated (up to 5.0 cm) using a pre-crosslinked hybrid. Scaffolds were assessed for the ability to support the growth of Escherichia coli using the Most Probable Number technique to quantify bacteria immediately after inoculation and 24 h later. This pre-crosslinking-based rheological property controlling technique can open a new avenue for 3D bio-fabrication of scaffolds, ensuring proper geometry. 

The promising success of 3D printing technique with synthetic polymers like nylon, ABS, PLA and epoxy motivates the researchers to put efforts into fabricating constructs with biocompatible natural polymers. The efforts have been broadened into various fields such as bioengineering, manufacturing, and regenerative medicine. Additive biomanufacturing commonly known as 3D bioprinting shows a lot of potential in tissue engineering with those natural polymers. Some challenges such as achieving printability, maintaining geometry in post printing stage, comforting encapsulated cells, and ensuring high proliferation are to be resolved to turn this process into a successful trial. Appropriate design of experiments with a detail rheological investigation can identify useful mechanical properties which is directly related to shape fidelity of 3D bio-printed scaffolds. As candidate natural polymers, Alginate-low viscous Carboxymethyl Cellulose (CMC) was used restricting the solid content 8% (w/v). Various rheological tests, such as the Steady Rate Sweep Test, Thixotropic (3ITT), Amplitude, and Frequency test were performed. The result indicated that rheological properties are CMC dependent. Printability and shape fidelity were analyzed of the filaments and scaffolds fabricated with all the combinations. The rheological results were co-related with the printability and shape fidelity result.